tbuckley/cabot-search huggingface dataset:
Abstract 1008: Cell density-related variability in chemotherapeutic resistance patterns in human cancer cells
Authors: Thomas Brown, Ujwal Punyamurtula, Jill Strandberg, Wafik S. El-Deiry
Journal: Cancer Research (2022)

Abstract Assessment of chemosensitivity and resistance varies with cancer cell of origin, drug and culture conditions. However, it is also not uncommon to find multiple different IC50 values for the same chemotherapeutic treatments acting on the same tumor cell lines. The aim of this research was to investigate this discrepancy in phenotypes and elucidate relevant variables that influence them. We assessed chemosensivity as a function of tumor cell density by plating TOV-21G ovarian, HT29 colorectal, and U2OS osteosarcoma cell lines in 96-well plates at various seeding densities per well and treated them with various concentrations of cisplatin, 5-fluorouracil (5-FU), and etoposide, respectively. CellTiter Glo assays were performed at 72 hours of treatment to determine IC50. Cisplatin IC50 for TOV-21G cells plated at 50,000, 20,000, 10,000, 5,000, 2,000, 1,000, and 500 cells/well were 12.00 μM, 5.47 μM, 3.16 μM, 2.45 μM, 2.54 μM, 2.83 μM, and 2.66 μM, respectively. Etoposide IC50 for U2OS cells plated at the same seeding densities were 6.50 μM, 5.19 μM, 3.23 μM, 2.98 μM, 2.49 μM, 2.13 μM, and 1.88 μM, respectively. IC50 values for HT29 cells treated with 5-FU at 2,000, 1,000, and 500 cells/well were 35.89 μM, 19.74 μM, and 13.42 μM, respectively. TOV-21G, U2OS, and HT29 cells all showed differences in IC50 values depending on cell plating density. TOV-21G and U2OS cells both showed a gradual decrease and plateau in IC50 as the cancer cell density decreased from 50,000 cells/well to 500 cells/well. HT29 cells displayed a sharper IC50 decrease as the cell density decreased from 2,000 cells/well to 500 cells/well. Our experiments demonstrate that IC50 values are not static at 72 hours and that tumor cell density is an important variable in influencing observed tumor resistance to traditionally effective drugs. As the density of cancer cells increases, the cells develop more resistance to the associated chemotherapy and require higher quantities of drug to achieve the same level of growth inhibition. We are currently exploring underlying molecular mechanisms that may influence these changes in IC50 including cell survival signaling proteins, cell-cell contact or growth factors that may influence cell density-related chemoresistance. Citation Format: Thomas Brown, Ujwal Punyamurtula, Jill Strandberg, Wafik S. El-Deiry. Cell density-related variability in chemotherapeutic resistance patterns in human cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1008.
Abstract 1885: Semi and fully automated immunostaining sample preparation platforms improve live leukocyte recovery, reproducibility, and cytometry data quality
Authors: Melvin Lye, Christoph Eberle, Ann Wang, Geoffrey K. Feld, Namyong Kim
Journal: Cancer Research (2022)

Abstract Limited innovation in automated cell and organelle sample preparation methodology limits the effectiveness of modern analytical methods, such as single-cell ‘omics, flow and mass cytometry. These techniques traditionally rely on manual centrifugation-based protocols for cell washing and suspension preparation, hampering researchers’ access to the reproducibility and scalability benefits of automation. We have developed a suite of cell suspension preparation systems that enable semi and full automation of cell washing protocols. These Laminar Wash࣪ technologies robustly, gently, and efficiently remove debris, dead cells, and unbound reagent using laminar flow and liquid handling robotics, rather than turbulent and harsh pelleting-plus-pipetting methods. Adaptation of standard protocols to Laminar Wash automation typically improves repetitive immunostaining processes and workflows, in terms of reduced hands-on time and inter- and intra-operator variability. We demonstrate the superior live cell retention and reproducibility of Laminar Wash over centrifugation in processing murine and humanized mouse peripheral blood mononuclear cells (PBMCs) and tumor infiltrating lymphocytes (TILs) for flow cytometry. Furthermore, we show how Laminar Wash improves flow cytometry data quality, in terms of debris removal and separation of immune cell subsets for both PBMCs and TILs. Overall, these results show how Laminar Wash methodology assists in standardizing sample preparation for cytometric analysis, an important and unmet need in cancer immunotherapy discovery and manufacturing workflows. Citation Format: Melvin Lye, Christoph Eberle, Ann Wang, Geoffrey K. Feld, Namyong Kim. Semi and fully automated immunostaining sample preparation platforms improve live leukocyte recovery, reproducibility, and cytometry data quality [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1885.
Abstract 864: Ophiopogonin D increase apoptosis by activating p53 via ribosomal protein L5 and L11 and inhibiting the expression of c-Myc via CNOT2
Authors: Hyun Min Ko, Wona Jee, Do-il Park, Somi Park, Ye-Rin Park, Hyeung-Jin Jang, Ji Hoon Jung
Journal: Cancer Research (2022)

Abstract Extracted from the root tuber of Ophiopogon japonicas, ophiopogon is well known to have an anti-cancer effect. However, the underlying mechanisms are still largely unknown. Here, we report that Ophiopogon D (OP-D) can inhibit colon cancer cell proliferation and induce apoptosis by inhibiting c-Myc expression through activation of p53 and CNOT2 regulation. Our results showed that OP-D induced p53 expression via ribosomal protein L5 or L11 and inhibited c-Myc expression through CNOT2 in a dose-dependent manner. Additionally, OP-D regulated cyclin D1 and CDK4 which are well known as cell cycle regulatory proteins. Consistently, OP-D inhibited the phosphorylation of AKT expression in a dose-dependent manner. Furthermore, OP-D shortened c-Myc’s half-life in a time-dependent manner. Furthermore, CNOT2 knockdown enhanced the inhibitory effect of OP-D on c-Myc in colon cancer cells. Interestingly, OP-D has increased the apoptotic effect of colon cancer cells when combined with doxorubicin or 5-FU, a treatment already used clinically. Altogether, our results suggested that OP-D regulates colon cancer cell survival and induces apoptosis by inhibiting c-Myc expression via activation of p53 and CNOT2 regulation. Citation Format: Hyun Min Ko, Wona Jee, Do-il Park, Somi Park, Ye-Rin Park, Hyeung-Jin Jang, Ji Hoon Jung. Ophiopogonin D increase apoptosis by activating p53 via ribosomal protein L5 and L11 and inhibiting the expression of c-Myc via CNOT2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 864.
Abstract LB190: Modulation of SPI1 transcriptional program contributes to the preclinical anti-tumor activity of SMARCA4/SMARCA2 ATPase inhibitors in AML
Authors: Gabriel J. Sandoval, Katharine Feldman, Sal Topal, Ammar Adam, Hsin-Jung Wu, Darshan Sappal, Luis M. Soares, David L. Lahr, Lan Xu, Rishi G. Vaswani, Jordana Muwanguzi, Liyue Huang, Jessica Piel, Mike Collins, Ho Man Chan, Michael J. Thomenius, Steven F. Bellon, Ryan G. Kruger, Carl P. Decicco, Richard C. Centore, Martin Hentemann
Journal: Cancer Research (2022)

Abstract SPI1 (PU.1) is an ETS family transcription factor that plays a critical role in hematopoietic development and differentiation. Regulation of SPI1 expression has also been implicated in Acute Myeloid Leukemia (AML) oncogenesis, though its mechanism is incompletely understood. Previously we have identified and characterized a series of novel dual inhibitors of the SMARCA4/SMARCA2 ATPases (also referred to as BRG1/BRM), critical components of the BAF (mSWI/SNF) family of chromatin remodeling complexes; and FHD-286, a related SMARCA4/SMARCA2 ATPase inhibitor, is currently being explored clinically for the treatment of metastatic uveal melanoma and AML (NCT04879017 and NCT04891757). Here we show that AML cell lines are sensitive to BAF ATPase inhibition, resulting in both cell cycle arrest and apoptosis. We demonstrate that BAF ATPase inhibition primarily affects the SPI1 transcriptional profile by regulating SPI1 genomic occupancy at various enhancer elements, resulting in downregulation of key target genes. Using in vivo mouse models of AML, we demonstrate dose-dependent tumor growth inhibition and pharmacodynamic modulation of SPI1 target genes. Together, these data suggest that modulation of SPI1 function may, in part, provide a mechanistic basis for the clinical development of FHD-286 in AML. Citation Format: Gabriel J. Sandoval, Katharine Feldman, Sal Topal, Ammar Adam, Hsin-Jung Wu, Darshan Sappal, Luis M. Soares, David L. Lahr, Lan Xu, Rishi G. Vaswani, Jordana Muwanguzi, Liyue Huang, Jessica Piel, Mike Collins, Ho Man Chan, Michael J. Thomenius, Steven F. Bellon, Ryan G. Kruger, Carl P. Decicco, Richard C. Centore, Martin Hentemann. Modulation of SPI1 transcriptional program contributes to the preclinical anti-tumor activity of SMARCA4/SMARCA2 ATPase inhibitors in AML [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB190.
Abstract 1495: Bioengineered RNAi molecules for cancer research and experimental therapy
Authors: Ai-Ming Yu
Journal: Cancer Research (2022)

Abstract RNAs have emerged as a novel group of medications, with distinct chemistry and pharmacology from conventional small molecules and proteins or antibodies. Indeed, some RNAi products (e.g., siRNAs) have been approved for clinical practice by the United State Food and Drug Administration while many others, including genome-derived miRNAs, are under active investigations. However, current research and development relies predominantly on the use of chemo-engineered RNA mimics made in vitro that are different from natural RNA molecules produced and folded in vivo. This is also in sharp contrast to protein research and therapy that primarily uses bioengineered or recombinant proteins produced and folded in vivo, instead of synthetic polypeptides or proteins made in vitro. Therefore, we have devoted large efforts to producing true biologic RNA molecules to better recapitulate the properties of natural RNA molecules for research and drug development. Herein I will present our one-of-a-kind RNA bioengineering technology that achieves high-yield, large-scale and cost-effective, in vivo fermentation production of biologic RNAs (BioRNAs) consisting of target RNAi molecules (e.g., miRNAs or siRNAs). Examples will be presented to demonstrate the precise release of payload RNAi molecules from BioRNAs in human cells and subsequently, modulate target gene expression and various cancer cellular processes. In addition, biologic RNAi prodrugs are effective to control tumor growth and metastasis in clinically relevant xenograft mouse models. Our results demonstrate that bioengineered RNA molecules are a novel class of agents for cancer research and drug development. Acknowledgements: This study was supported by grants R01CA225958, R01CA253230 and R35GM140835 from NIH. Citation Format: Ai-Ming Yu. Bioengineered RNAi molecules for cancer research and experimental therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1495.
Abstract 2117: IFN-γ signaling in myeloid cells regulates pancreatic cancer growth and progression
Authors: Elena Ivleva, Natalia Andreeva, Sergei Grivennikov
Journal: Cancer Research (2022)

Abstract Pancreatic ductal adenocarcinoma (PDAC) is characterized by low survival, early metastasis, and rapidly emerging resistance to therapy. Interferon gamma (IFN-γ) signaling pathway is one of the key regulators of the tumor microenvironment in numerous cancer settings and its possible key role in myeloid cells is suggested. Here we investigated the IFN-y signaling in myeloid cells during PDAC development, progression, and metastasis using mice with conditional ablation of IFNgR2 in myeloid cells (IFNgR2-Delta-Mye). We performed an orthotopical injection of cells derived from conditional murine pancreatic model (Pdx1Cre-Kras-P53, or “KPC” cells) into IFNgR2-LysMCre (IFNgR2-Delta-Mye) mice. Ablation of IFN-γ signaling in myeloid cells led to a higher tumor burden compared to controls. Subsequent RNA sequence analysis demonstrated that IFN-γR2 deficiency in myeloid cells resulted in increased TGFβ1 gene expression as well as changes in metabolic and growth factor pathways in macrophages isolated from tumor tissue. Immunofluorescent analysis demonstrated that IFN-γR2 deletion in myeloid cells led to changes in cancer cell differentiation, resulting in a loss of characteristic epithelial histological pattern and shifting it towards more aggressive phenotype, possible via TGFβ1 dependent mechanism. Using murine model of liver metastasis, we found that loss of IFN-γR2 in myeloid cells leads to increased metastatic outgrowth of PDAC KPC cells. This effect was partially dependent on the gut microbiota, as shown by “separated and co-housed mice” experiments. Deficiency of IFN-γR2 in myeloid cells resulted in increased recruitment and infiltration of monocytes in normal and tumor/metastatic liver tissues. Our data identifies myeloid cell type specific IFN-γ signaling pathway as an important regulator of pancreatic cancer progression and metastasis, that can be potentially exploited as a novel immunooncology target. Citation Format: Elena Ivleva, Natalia Andreeva, Sergei Grivennikov. IFN-γ signaling in myeloid cells regulates pancreatic cancer growth and progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2117.
Abstract 2180: Deciphering the Warburg effect: Redox is the key to tumor differentiation
Authors: Jiang Haowen, Rachel L. Greathouse, Bo He, Yang Li, Albert M. LI, Balint Forgo, Selene Banuelos, Joshua Gruber, Hiroyuki Shimada, Bill Chiu, Jiangbin Ye
Journal: Cancer Research (2022)

Abstract It was proposed by Warburg that the cause and consequence of the Warburg effect were the injury of respiration and cell dedifferentiation, respectively. However, the underlying mechanism was unclear due to limited understanding of metabolic control of epigenetics. Inhibition of mitochondrial electron transport chain (ETC) causes reduction of NAD+/NADH ratio. Recently it was discovered that this redox stress not only drives lactate production from pyruvate to enhance the Warburg effect, but also promotes the conversion of α-ketoglutarate (αKG) to 2-hydroxyglutarate (2HG), turning the substrate of DNA demethylase TET into an inhibitor of TET, suggesting a potential link between ETC inhibition and dysregulated DNA methylation in cancer. In this study we use a mitochondrial uncoupler niclosamide ethanolamine (NEN) to block the Warburg effect and remodel cancer epigenome. NEN treatment activates ETC and accelerates glutaminolysis to produce αKG, which promotes promoter CpG island demethylation. This epigenomic reprograming not only upregulates tumor suppressor genes such as p53, inhibits N-Myc expression, but also activates neuron differentiation markers to promote differentiation in neuroblastoma cells. Hypoxia is one common microenvironmental stress existing in all solid tumor. NEN treatment reduces 2HG generation from αKG under hypoxia, highlighting its application potential in preventing DNA hypermethylation in vivo. Associated with 2HG reduction, NEN treatment also represses HIF signaling under hypoxia. In addition, NEN treatment inhibits reductive carboxylation flux. Moreover, dietary supplementation with NEN reduces orthotopic neuroblastoma growth and N-Myc expression in vivo. Together, these results validated Warburg’s hypothesis that mitochondrial respiration is the key to maintain normal differentiation. Restoring ETC activity with mitochondrial uncoupler is an effective metabolic/epigenetic therapy against neuroblastoma (Jiang et al., BioRxiv 2021). Citation Format: Jiang Haowen, Rachel L. Greathouse, Bo He, Yang Li, Albert M. LI, Balint Forgo, Selene Banuelos, Joshua Gruber, Hiroyuki Shimada, Bill Chiu, Jiangbin Ye. Deciphering the Warburg effect: Redox is the key to tumor differentiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2180.
Abstract 1098: Structure-activity relationship of pentacyclic triterpenes against chemoresistance and metastasis on non-small lung adenocarcinoma cells
Authors: Yamixa Delgado, Daraishka Perez, Grace Torres
Journal: Cancer Research (2022)

Abstract Lung cancer is the second most common and leading cause of cancer-related death worldwide, 85% of these cases being non-small cell lung carcinoma (NSCLC). Although chemotherapy is the first-line treatment for lung cancer, its effectiveness is hampered by the dose-limiting toxic effects, chemoresistance, and metastatic process. Recently, plant-derived cyclic triterpenoids (eg, cholesterol-structure like compounds) have shown a wide spectrum of pharmacological activities, especially their potential against different cancer cells. Herein, we investigated the structure-activity relationship of several pentacyclic triterpenes: oleanolic acid (OleA), ursolic acid (UrA), betulinic acid (BeA), Asiatic acid (AsA), betulin (BeN), lupeol (Lupe), diosgenin (Dio), stigmasterol (Stg) and β-sitosterol (β-Sito) on NSCLC A549 cells. From our studies, the IC50 for OleA, UrA, BeA, AsA, BeN, Lupe, Dio were in the low µM range (from 10-75 µM) after 24h of incubation. β-Sito and Stg did not show any significant cytotoxic pattern in this µM range. In addition, BeA showed the highest cytotoxic pattern in these cells. We performed metabolic activity assays to determine the mechanism of action of these triterpenes. Mitochondrial membrane and caspases activity assays showed that all the cytotoxic triterpenes induced activation of caspase-3 and decline in the mitochondrial potential, except BeA and AsA. Our results also show that all the cytotoxic triterpenes disrupt cell membrane, except for OleA. However, there is no reactive oxygen species production for all of them. Gene expression assays of chemoresistance- and metastasis-related genes (EGFR, VEGF, MMP9, Pgp, TP53, NDRG1, CHD4) using qPCR demonstrated the downregulation of all of these genes after the incubation with the cytotoxic triterpenes. These results demonstrated that even slight structural changes in these triterpenes could influence the cytotoxic and metabolic pathways responses. This study opens promising perspectives for further research on the role of phytochemical triterpenes, which ultimately will contribute to a more rational application in lung cancer therapy. Citation Format: Yamixa Delgado, Daraishka Perez, Grace Torres. Structure-activity relationship of pentacyclic triterpenes against chemoresistance and metastasis on non-small lung adenocarcinoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1098.
Collaborator Name Misspelled
Authors: Keith Couper, Bronwen Connolly, Daniel Mcauley
Journal: JAMA (2022)

and other sequalae associated with invasive mechanical ventilation, avoidance of tracheal intubation is itself an important outcome to patients, clinicians, and health systems. 3,4 The pragmatic nature of the trial and the context in which it was delivered precluded collection of granular physiological data in patients who did not undergo intubation.We agree with the final point made by Yarnell and Sklar that the optimum time point for tracheal intubation remains unclear. 5,6It was in part for this reason that we decided not to protocolize decisions relating to the timing of tracheal intubation.The decision to undertake tracheal intubation for most patients encompasses a range of factors including current physiology, subjective respiratory assessment (eg, work of breathing), trajectory of illness, and patient wishes, which are difficult to operationalize in the trial setting.
Abstract 5476: Novel azetidine-based STAT3 inhibitors induce misfolded protein response, endoplasmic reticulum stress and mitophagy, and inhibit breast tumor growth <i>in vivo</i>
Authors: Peibin Yue, Francisco Lopez-Tapia, Yinsong Zhu, Nagendra Verma, Felix Alonso-Valenteen, Simoun Mikhael, Lali Medina-Kauwe, Marcus Tius, James Turkson
Journal: Cancer Research (2022)

Abstract Anticancer drugs that directly target Signal Transducer and Activator of Transcription (Stat)3 have remained elusive. We present azetidine-based small molecules as a new class of potent Stat3 inhibitors. Compounds H172 and H182 have potency (IC50) of 0.38-0.98 µM, H105 and H120 have IC50 of 1.75-2.07 µM against Stat3 DNA-binding activity in vitro, compared to the IC50 of &amp;gt;17 µM against Stat1 or Stat5 activity. Treatment of the triple-negative breast cancer (TNBC), MDA-MB-231 and MDA-MB-468 cells with the new compounds inhibited constitutive Stat3 activation, promoted Stat3 ubiquitination, blocked anchorage-dependent and independent growth, with potency, EC50 of 1.0-1.9 μM, and induced apoptosis of these cells. By contrast, pYStat1, pYStat5, pS727Stat3, pSrc, pJanus kinase, or the induction of other signaling events are not inhibited in compound-treated cells, and normal or tumor cells that do not harbor aberrantly-active Stat3 are only weakly inhibited. In vivo delivery of H120 or H182 as a single agent inhibited growth of human TNBC xenografts, and of H278 (HCl salt of H182) combined with radiation abrogated tumor growth and improved survival in mouse TNBC syngeneic models. These studies provide novel mechanism of disruption of aberrantly-active Stat3 and its dynamics in showing that the azetidine-based Stat3 inhibitors trigger Stat3 ubiquitination, and tumor cell death. H182 and H172 are new entities for further therapeutic development against TNBC and other cancers that harbor aberrantly-active Stat3. Citation Format: Peibin Yue, Francisco Lopez-Tapia, Yinsong Zhu, Nagendra Verma, Felix Alonso-Valenteen, Simoun Mikhael, Lali Medina-Kauwe, Marcus Tius, James Turkson. Novel azetidine-based STAT3 inhibitors induce misfolded protein response, endoplasmic reticulum stress and mitophagy, and inhibit breast tumor growth in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5476.